Irurzun A, Nieva J L, Carrasco L
Centro de Biologia Molecular, UAM-CSIC, Universidad Autonoma de Madrid, Canto Blanco, Spain.
Virology. 1997 Jan 20;227(2):488-92. doi: 10.1006/viro.1996.8340.
Semliki forest virus (SFV) was biosynthetically labeled with pyrene phospholipids and used to investigate two alternative routes of entry of SFV into BHK-21 cells: (1) receptor-mediated endocytosis followed by fusion of the viral envelope with the endosomal membrane and (2) direct fusion of SFV with the plasma membrane induced by low pH treatment. The selective inhibitor of the vacuolar proton-ATPase, concanamycin A, abolished fusion and subsequent infection only when the virus utilized the endocytic route to enter cells. The inhibitory effect of this macrolide antibiotic was bypassed by low pH treatment of cells. However, the ionophore nigericin was inhibitory irrespective of the route used by the virus to infect cells, suggesting the necessity of a transmembrane pH gradient for the entry process. According to our results, concanamycin A emerges as a suitable tool for selectively investigating the involvement of endosomal function in animal virus entry.
用芘磷脂对塞姆利基森林病毒（SFV）进行生物合成标记，并用于研究SFV进入BHK - 21细胞的两种替代途径：（1）受体介导的内吞作用，随后病毒包膜与内体膜融合；（2）低pH处理诱导SFV与质膜直接融合。液泡质子 - ATP酶的选择性抑制剂 concanamycin A，仅在病毒利用内吞途径进入细胞时才会消除融合及随后的感染。通过对细胞进行低pH处理可绕过这种大环内酯类抗生素的抑制作用。然而，离子载体尼日利亚菌素无论病毒感染细胞所采用的途径如何均具有抑制作用，这表明跨膜pH梯度对于进入过程是必需的。根据我们的结果，concanamycin A成为选择性研究内体功能在动物病毒进入过程中所起作用的合适工具。
